Objective. Toothbrush abrasion is significant in the development of tooth wear, particularly when combined with erosion. This in vitro study aimed to evaluate the impact of toothpaste slurry abrasivity and toothbrush filament stiffness on abrasion of eroded enamel. Material and methods. Eroded enamel samples (hydrochloric acid, pH: 2.6, 15 s) were brushed with 40 strokes in an automatic brushing machine using manual toothbrushes with different filament stiffness (filament diameter: 0.15, 0.20, or 0.25 mm). A paste-free control slurry (relative enamel abrasion (REA) value 2) and toothpaste slurries with different abrasivity (REA values 6 or 9) were used for brushing. Erosion and abrasion were followed by storing the enamel samples in artificial saliva for 3 h. After each 4th cycle, the samples were stored in artificial saliva for 15 h. After 60 cycles, enamel loss was measured by profilometry and statistically analyzed by two-way and one-way ANOVA and Bonferroni/Dunn post-hoc tests. Results. Loss of enamel (mean, µm) was influenced mainly by the abrasivity of the slurry and increased along with REA value (REA 2: 0.0-0.2, REA 6: 2.1-3.3, REA 9: 2.9-3.7). Abrasion of eroded enamel was also affected by filament stiffness of the toothbrush, but only groups brushed with toothpaste slurry of REA 6 showed any significant difference between the different toothbrushes. Thereby, toothbrushes with 0.2 mm filament diameter caused higher enamel loss than 0.15 and 0.25 mm filaments. Conclusions. Toothbrush abrasion of eroded enamel is influenced mainly by the abrasivity of the toothpaste slurry, but is also modified by toothbrush filament stiffness.